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To face this problem, we set up a CRISPR\/Cas9\u2010based strategy for knock\u2010in cellular model generation, focusing on inherited metabolic disorders (IMDs). We selected variants in seven IMD\u2010associated genes, including seven reported disease\u2010causing variants and four benign\/likely benign variants. Overall, 11 knock\u2010in cell models were generated via homology\u2010directed repair in HAP1 haploid cells using CRISPR\/Cas9. The functional impact of the variants was determined by analyzing the characteristic biochemical alterations of each disorder. Functional studies performed in knock\u2010in cell models showed that our approach accurately distinguished the functional effect of pathogenic from non\u2010pathogenic variants in a reliable manner in a wide range of IMDs. Our study provides a generic approach to assess the functional impact of genetic variants to improve IMD diagnosis and this tool could emerge as a promising alternative to invasive tests, such as muscular or skin biopsies. Although the study has been performed only in IMDs, this strategy is generic and could be applied to other genetic disorders.<\/jats:p>","DOI":"10.1002\/jimd.12681","type":"journal-article","created":{"date-parts":[[2023,9,18]],"date-time":"2023-09-18T03:31:58Z","timestamp":1695007918000},"page":"1029-1042","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["<scp>CRISPR<\/scp>\n                    \/Cas9\u2010based functional genomics strategy to decipher the pathogenicity of genetic variants in inherited metabolic disorders"],"prefix":"10.1002","volume":"46","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6863-3993","authenticated-orcid":false,"given":"Gerard","family":"Mu\u00f1oz\u2010Pujol","sequence":"first","affiliation":[{"name":"Secci\u00f3 d'Errors Cong\u00e8nits del Metabolisme\u2010IBC, Servei de Bioqu\u00edmica i Gen\u00e8tica Molecular Hospital Cl\u00ednic de Barcelona, IDIBAPS, CIBERER  Barcelona 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